Hayat cp3

Kingdom of Saudi Arabia
Taibah University
Collage of applied medical sciences
Department of Clinical Laboratory Sciences
GeneXpert Medical Laboratory

technique
COURSE TITLE:
CLINICAL PRACTICE 3 ( MLT 465)
COURSE COORDINATOR:
DR. HANAN ALRASHIDI
STUDENT NAME:
HAYAT MARWAN KUWAITI
STUDENT ID: 4158675
[email protected]
WORD COUNT:459
SUBMISSION DATE: NOVEMBER 2023
Introduction
Gene-Xpert, a CBNAAT (catridge-based nucleic acid amplification test), is a commonly used

Tuberculosis diagnostic test. This is a fast diagnostic test for tuberculosis identification as well as
Rifampicin resistance in circumstances where a direct smear is negative. It is still uncommon in
developing nations such as India due to a lack of availability and cost effectiveness.
Principle
The Xpert MTB/RIF test for use with the Cepheid GeneXpert® System is a semi-quantitative nested
real-time PCR in vitro diagnostic test for: 1) the detection of Mycobacterium tuberculosis complex
(MTB complex) DNA in sputum samples or concentrated sediments prepared from induced or
expectorated sputa that are either acid-fast bacilli (AFB) smear positive or negative
Procedure
1. Collection and Transportation of Specimens
• Collect at least 1 mL of sputum per specimen. The subject must be either seated or standing.
• Whenever possible, specimens should be stored at 2-8 °C prior to processing.
However, the specimens can be stored at a maximum of 35 °C for 3 days and then at 2-8 °C for an
additional 7 days if necessary.
1) Rinse the patient's mouth with water twice.
2) Remove the sputum collection container's lid.
3) Instruct the patient to take a deep breath, cough vigorously, and expectorate the material into the
sterile screw-capped specimen collection container. Avoid spills and soiling the container's outside.
4) Close the collection device's cover.
5) When possible, specimens should be kept at 2-8 °C, including throughout transport.
2. Sputum Expectorated Samples
1
3. Sputum Sediments
• Accept only as many samples as there are modules available to execute the test on the GeneXpert Dx
System at any given time. Sputum sediments generated following the Kent and Kubica method and
resuspended in a 67mM Phosphate/H2O buffer can be assayed using Xpert MTB/RIF. When the re-
suspension is ready for normal laboratory smear or culture tests, make sure there is at least 0.5 mL of re-
suspended sediment available to run Xpert MTB/RIF.
1) Write the sample ID on each Xpert MTB/RIF cartridge. (Write on the cartridge's sides or place an ID
label.) Note: Do not place the label on the cartridge's lid or impede the cartridge's current 2D barcode.
2) Using a sterile transfer pipette, transfer at least 0.5 mL of the whole re-suspension pellet to a conical,
screw-capped tube for the Xpert MTB/RIF.
The complete sample can also be processed in the original tube.
3) If re-suspended sediments are not immediately processed for Xpert MTB/RIF, store them at 2-8 °C.
Store for no more than 12 hours.
4) Using a sterile transfer, add 1.5 mL of Xpert MTB/RIF Sample Reagent (SR) to 0.5 mL of re-
suspension sediment sample.
5) Shake 10 - 20 times forcefully. One back-and-forth movement equals one shaking.
6) Incubate the specimen at room temperature for 15 minutes. Shake the specimen forcefully 10 - 20 times
more between 5 and 10 minutes of incubation. There should be no apparent clumps of sputum in the
samples. Particulate matter that is not part of the sample may exist.
4. Preparing the Cartridge
5. Starting the Test
Advantages
The main benefits of the test for diagnosis are its reliability when compared to sputum microscopy and its
speed of results when compared to the culture test. Although sputum microscopy is quick and inexpensive
for diagnosing tuberculosis, it is frequently unreliable. It is especially untrustworthy when persons are HIV
positive. Although culture provides a reliable diagnosis, the process normally takes weeks rather than the
hours required by the Genexpert test.
The biggest advantage in finding rifampicin resistance is, once again, speed. Normally, it takes weeks
rather than hours to obtain a medication resistance result.
2
Limitation
The Xpert MTB/RIF's performance was validated using the techniques outlined in this package insert.
Changes to these methods may affect the test's performance. The Xpert MTB/RIF results should be
analyzed in conjunction with the clinician's other laboratory and clinical findings.
Because MTB detection is based on the amount of organisms present in the sample, accurate results are
contingent on adequate specimen collection, handling, and storage.
Improper samples may result in incorrect test findings, collection, failure to follow the appropriate sample
collection process, handling or storage, technical error, sample mix-up, or insufficient starting material
concentration. To avoid incorrect results, carefully follow the instructions in this insert.
A positive test result does not always imply the presence of live organisms. However, the presence of MTB
and Rifampicin resistance is presumed.
Prior or concurrent antibiotic therapy may have an impact on test results.
As a result, therapeutic success or failure cannot be determined using this test because DNA may survive
after antimicrobial therapy.
Mutations or polymorphisms in primer or probe binding areas may interfere with detection of novel or
unknown MDR-MTB or rifampicin resistant strains, leading to a false negative result.
Application
The WHO advised that the test be used as the first line of defense in anyone suspected of having MDR TB
or HIV-associated TB. They also proposed that, due to the lack of accuracy of smear microscopy, it may be
used as a follow-up test to microscopy in settings where MDR TB and/or HIV are of less concern,
particularly in smear negative specimens. They did, however, acknowledge the significant resource
consequences of employing it in this second manner.
Conclusion
The GeneXpert assay is a novel method for the rapid identification of MTB and treatment resistance. It is
an appealing point-of-care test, with high sensitivity and specificity and a turnaround time of 2 hours,
allowing for rapid diagnosis and proper management of tuberculosis pleuritis and pericarditis.
3
References
1. Genexpert - testing for TB & Drug Resistant TB [Internet]. 2022 [cited
2023 Nov 14]. Available from: https://tbfacts.org/genexpert/
2. [Internet]. [cited 2023 Nov 14]. Available from:

https://www.moleculartb.org/files/documents/2
3. Saeed M, Ahmad M, Iram S, Riaz S, Akhtar M, Aslam M.

GeneXpert technology. A breakthrough for the diagnosis of
tuberculous pericarditis and pleuritis in less than 2 hours [Internet].
U.S. National Library of Medicine; 2017 [cited 2023 Nov 15]. Available
from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5556276/
4. Prakash AK, Datta B, Goyal P, Chatterjee P, Gupta G. Gene-xpert

gives early diagnosis in early tuberculosis [Internet]. European
Respiratory Society; 2016 [cited 2023 Nov 15]. Available from:
https://erj.ersjournals.com/content/48/suppl_60/PA2775

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Kingdom of Saudi Arabia

GeneXpert Medical Laboratory

Gene-Xpert, a CBNAAT (catridge-based nucleic acid amplification test), is a commonly used

1. Collection and Transportation of Specimens

2. Sputum Expectorated Samples

4. Preparing the Cartridge

5. Starting the Test

2. [Internet]. [cited 2023 Nov 14]. Available from:

3. Saeed M, Ahmad M, Iram S, Riaz S, Akhtar M, Aslam M.

4. Prakash AK, Datta B, Goyal P, Chatterjee P, Gupta G. Gene-xpert

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